title: Rescue of maturation off-pathway products in the assembly of Pseudomonas phage φ 6.
abstract: Many complex viruses use an assembly pathway in which their genome is packaged into an empty procapsid which subsequently matures into its final expanded form. We utilized Pseudomonas phage 6, a well-established virus assembly model, to probe the plasticity of the procapsid maturation pathway. The 6 packaging nucleoside triphosphatase (NTPase), which powers sequential translocation of the three viral genomic single-stranded RNA molecules to the procapsid during capsid maturation, is part of the mature 6 virion but may spontaneously be dissociated from the procapsid shell. We demonstrate that the dissociation of NTPase subunits results in premature capsid expansion, which is detected as a change in the sedimentation velocity and as defects in RNA packaging and transcription activity. However, this dead-end conformation of the procapsids was rescued by the addition of purified NTPase hexamers, which efficiently associated on the NTPase-deficient particles and subsequently drove their contraction to the compact naive conformation. The resulting particles regained their biological and enzymatic activities, directing them into a productive maturation pathway. These observations imply that the maturation pathways of complex viruses may contain reversible steps that allow the rescue of the off-pathway conformation in an overall unidirectional virion assembly pathway. Furthermore, we provide direct experimental evidence that particles which have different physical properties (distinct sedimentation velocities and conformations) display different stages of the genome packaging program and show that the transcriptional activity of the 6 procapsids correlates with the number of associated NTPase subunits.
authors.affiliation: Department of Biosciences.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Sun
authors.forename: Markus J
authors.initials: MJ
authors.lastname: Pirttimaa
authors.forename: Dennis H
authors.initials: DH
authors.lastname: Bamford
authors.forename: Minna M
authors.initials: MM
authors.lastname: Poranen
mesh_terms: D017126:Bacteriophage phi 6 / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D044224:Pseudomonas syringae / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins; D014764:Viral Proteins